Project Summary/Abstract: The proposed project is a high-risk, high-impact effort to achieve the early steps demonstrating the use of low-temperature corona discharge generated inside the cardiovascular system as a less-invasive treatment of infectious bacterial biofilm growths on native tissue and implanted medical devices. The long-term impact of the successful project is the less-invasive treatment of biofilm infections such as infective endocarditis (IE), which has a 30% mortality rate at 1 year, through a catheterized low-temperature plasma (LTP) device to produce localized doses of reactive oxygen species (ROS) in the vicinity of the bacterial colony. ROS produced through gas-phase LTP has been demonstrated to destroy bacteria associated with IE, including staphylococcus aureus, as well as bacterial biofilms. This is part of an exciting, emerging field known as plasma medicine, which, to date, is focused on external and in-vitro applications. This project will open up a new and rich area of investigation for plasma medicine, internal plasma medicine, which has the potential to impact a wide range of disease states in the human body through less invasive means than surgery or systemic drugs. The project consists of three Specific Aims to evaluate the effect of LTP generation in blood, the effects on bacterial cultures in the vicinity, and the miniaturization of the LTP system for future integration with catheters. The first Aim is to determine the effect of LTP generation in human whole blood on the blood constituents' structure and function and measurement of ROS generation in the liquid part of the blood. The second Aim applies the most favorable LTP conditions to bacterial cultures (S. aureus, P. aeruginosa and S. pneumoniae) in a blood environment and evaluates the destructive effects of the ROS generation on unprotected cultures and biofilm specimens. The third Aim will demonstrate the potential for future translation of these devices to clinical practice by miniaturizing them to fit on the end of a catheter. Successful completion of this project will open the door to wider investigations under the R01 mechanism of the effect of LTP generation on healthy adjacent tissue, understanding the transport of LTP-generated ROS in the cardiovascular system, and investigations of other internal disease states.